An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.
Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Tissue - Human Salivary glands
As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.
As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.
Get tips on using miRcute miRNA Isolation Kit to perform RNA isolation / purification Cells - immortalized MDA-MB-157
Get tips on using TaqMan® MicroRNA Reverse Transcription Kit to perform siRNA / RNAi /miRNA transfection Mouse - Glomerular Mesangial cells polymer / lipid
Get tips on using Mammary Epithelial Cell Growth Medium to perform 3D Cell Culture Media Human primary breast ephitelial cells-organoids
Get tips on using Mammary Epithelial Cell Growth Medium to perform 3D Cell Culture Media Human primary breast ephitelial cells-Mammospheres
Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary hematopoietic stem cells
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